LOCUS       V044670                 9390 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044670
VERSION     V044670
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             234..414
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    458..583
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1076..1309
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1493..1537
                     /codon_start=1
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /product="antigenic peptide corresponding to amino acids
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
                     et al., 2013)"
                     /transl_table=1
                     /translation="KNEQELLELDKWASL"
                     /label="gp41 peptide"
     misc_feature    1798..1915
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2005..2208
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2297..3409
                     /label="Pcbp3(NM_021568)"
                     /note="Pcbp3(NM_021568)"
                     /gene="Pcbp3"
     CDS             3416..3505
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     promoter        3549..3760
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3773..4041
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4070..4079
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4100..4762
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /transl_table=1
                     /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
                     FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
                     EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
                     YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
                     /label="CopGFP"
     CDS             4832..4885
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from&nbsp;Thosea asigna virus capsid
                     protein"
                     /protein_id=""
                     /transl_table=1
                     /translation="EGRGSLLTCGDVEENPGP"
                     /label="T2A"
     CDS             4886..5485
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     misc_feature    5486..6073
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6147..6380
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6452..6573
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6592..6727
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6765..6781)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6789..6805
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(6813..6843)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6858..6879
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7167..7755)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7926..8786)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(8787..8891)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9365..9381
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggttaacttt
     1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga
     1921 tactagtatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     1981 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     2041 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     2101 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     2161 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
     2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagagc
     2281 tagcgaattc gccaccatgg gggaaggtga cgccatctgg gccccaccca tccttcctca
     2341 cagcaccctc ggcaccttaa gccaccaccc tgagctacat tttggaggga agatggaatc
     2401 taaggtctcg gaaggtggcc tgaacgtaac cctgaccatc cgcctactga tgcatggaaa
     2461 ggaagttgga agcatcatcg ggaagaaagg agaaacagta aagaagatgc gtgaagagag
     2521 tggcgccagg atcaacatct cagagggaaa ctgcccagag agaatcgtta caatcacagg
     2581 cccaactgac gccatcttca aggcctttgc tatgatcgcg tacaagtttg aggaggacat
     2641 cattaattcc atgagcaaca gccctgccac cagcaagcct ccagtgacac tgcgactagt
     2701 ggtccctgcc agccagtgtg ggtccctgat cggcaaaggt ggctccaaga tcaaggagat
     2761 ccgggagtcc acaggtgctc aggtccaggt ggcaggtgac atgctgccca actccacaga
     2821 gcgggcagtg actatctcag ggaccccaga cgccatcatc cagtgtgtga aacagatctg
     2881 tgtggtcatg ctggagtccc caccgaaagg tgccaccatt ccctaccgcc caaagcccgc
     2941 ctccacccct gtcatttttg caggtggtca ggcctacaca atccagggac agtacgccat
     3001 cccccaccca gatcagttga ccaagctcca ccagttggcc atgcagcaaa ccccctttcc
     3061 tcccctcgga cagaccaacc ccgctttccc cggagaaaag ctgcctttac actcctccga
     3121 agaagctcaa aatctgatgg gccagtcgtc aggtctggat gccagcccac cggccagcac
     3181 tcatgagctc accattccca atgatcttat aggctgcata attggacgcc aaggcaccaa
     3241 aatcaatgaa atccgacaga tgtctggagc tcagatcaaa atcgccaatg ccactgaagg
     3301 gtcgtcagag cgccagatta ccatcacagg gaccccagcc aacatcagcc ttgcccagta
     3361 tctcatcaac gctaggctga cgtctgaggt caccgggatg ggtgcactcg gatccgaaca
     3421 aaaactaata tcagaggaag atttggaaca gaaactaatc agtgaagaag atttagagca
     3481 aaagttaatt tccgaggagg acttatagcg gccgcgaagg atctgcgatc gctccggtgc
     3541 ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg
     3601 caattgaacg ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta
     3661 ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt
     3721 gaacgttctt tttcgcaacg ggtttgccgc cagaacacag ctgaagcttc gaggggctcg
     3781 catctctcct tcacgcgccc gccgccctac ctgaggccgc catccacgcc ggttgagtcg
     3841 cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg tccgccgtct aggtaagttt
     3901 aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct tggagcctac ctagactcag
     3961 ccggctctcc acgctttgcc tgaccctgct tgctcaactc tacgtctttg tttcgttttc
     4021 tgttctgcgc cgttacagat ccaagctgtg accggcgcct acgctagacg ccaccatgga
     4081 gagcgacgag agcggcctgc ccgccatgga gatcgagtgc cgcatcaccg gcaccctgaa
     4141 cggcgtggag ttcgagctgg tgggcggcgg agagggcacc cccaagcagg gccgcatgac
     4201 caacaagatg aagagcacca aaggcgccct gaccttcagc ccctacctgc tgagccacgt
     4261 gatgggctac ggcttctacc acttcggcac ctaccccagc ggctacgaga accccttcct
     4321 gcacgccatc aacaacggcg gctacaccaa cacccgcatc gagaagtacg aggacggcgg
     4381 cgtgctgcac gtgagcttca gctaccgcta cgaggccggc cgcgtgatcg gcgacttcaa
     4441 ggtggtgggc accggcttcc ccgaggacag cgtgatcttc accgacaaga tcatccgcag
     4501 caacgccacc gtggagcacc tgcaccccat gggcgataac gtgctggtgg gcagcttcgc
     4561 ccgcaccttc agcctgcgcg acggcggcta ctacagcttc gtggtggaca gccacatgca
     4621 cttcaagagc gccatccacc ccagcatcct gcagaacggg ggccccatgt tcgccttccg
     4681 ccgcgtggag gagctgcaca gcaacaccga gctgggcatc gtggagtacc agcacgcctt
     4741 caagaccccc atcgccttcg ccagatcccg cgctcagtcg tccaattctg ccgtggacgg
     4801 caccgccgga cccggctcca ccggatctcg cgagggcaga ggaagtcttc taacatgcgg
     4861 tgacgtggag gagaatcccg gccctatgac cgagtacaag cccacggtgc gcctcgccac
     4921 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     4981 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     5041 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     5101 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     5161 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     5221 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     5281 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     5341 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
     5401 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     5461 catgacccgc aagcccggtg cctgaaatca acctctggat tacaaaattt gtgaaagatt
     5521 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     5581 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     5641 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     5701 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     5761 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     5821 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     5881 gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     5941 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     6001 ggctctgcgg cctcttccgc gtctccgcct tcgccctcag acgagtcgga tctccctttg
     6061 gccgcctccc cgcctggtac ctttaagacc aatgacttac aaggcagctg tagatcttag
     6121 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaaaataaga
     6181 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
     6241 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     6301 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     6361 agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt atttataact
     6421 tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta
     6481 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     6541 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc
     6601 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     6661 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     6721 ggaggcctag acttttgcag agacggccca aattcgtaat catggtcata gctgtttcct
     6781 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     6841 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     6901 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     6961 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     7021 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     7081 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     7141 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     7201 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     7261 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     7321 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     7381 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     7441 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     7501 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     7561 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     7621 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     7681 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     7741 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     7801 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     7861 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     7921 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     7981 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     8041 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     8101 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     8161 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     8221 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     8281 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     8341 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     8401 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     8461 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     8521 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     8581 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     8641 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     8701 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     8761 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     8821 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     8881 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     8941 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     9001 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     9061 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     9121 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
     9181 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc gccattcagg
     9241 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     9301 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     9361 cgttgtaaaa cgacggccag tgccaagctg
//
